Bcl-2 expression in sputum eosinophils in patients with acute asthma

Eosinophils play a pivotal role in asthmatic airway inflammation. Apoptosis is thought to be critically relevant in promoting the clearance of inflammatory cells and the resolution of inflammation. Bcl-2 inhibits apoptosis in cellular systems. A study was undertaken to determine whether bcl-2 expression in sputum reflects the clinical severity of patients with asthma. The relationship between bcl-2 expression in sputum eosinophils and eosinophil activation markers such as interleukin (IL)-5 and eosinophil cationic protein (ECP) levels in sputum supernatant was evaluated.

Sputum was obtained from 18 patients with asthma. Fresh expectorated sputum separated from saliva was treated with an equal volume of dithiothreitol 0.1%, cytospun for cell differentials and bcl-2 stain, and the supernatant was collected for biochemical assay. Bcl-2+ eosinophils were stained using immunocytochemistry, ECP was measured by fluoroimmunoassay, and IL-5 was detected by sandwich enzyme linked immunosorbant assay.

Twelve patients with severe or life threatening asthma had more bcl-2+ eosinophils (mean difference 46.8% (95% CI 27.0 to 66.6), p<0.01) and a higher ECP level (p<0.01) in the sputum than those with mild to moderate asthma (n = 6). IL-5 was frequently detected in patients with severe or life threatening asthma (11/12 versus 1/6, p<0.01). There was a significant positive correlation between bcl-2+ eosinophils and ECP levels (r = 0.61, p<0.01) and between bcl-2+ eosinophils and IL-5 levels (r = 0.83, p<0.01). There was a significant negative correlation between bcl-2+ eosinophils and FEV(1)/FVC (r = -0.54, p<0.05).

The increased expression of bcl-2 in eosinophils from sputum of subjects with severe asthma suggests that bcl-2 may prolong survival and decrease apoptosis of airway eosinophils in asthma.